Tumor immunology BC Yang

1. Tumor immunologyBC Yang 100 60 10 0 year Cancer/ Neuronal disorder Infection/Development Expected life-span Immune capability 2010

2. 衛生署資料，台灣人十大死因 98年 癌症、心臟疾病（不含高血壓）、腦血管疾病、肺炎、糖尿病、事故傷害、慢性下呼吸道疾病、慢性肝病及肝硬化，自殺、腎臟疾病。

3. 1960 Nobel prize in physiology or Medicinefor discovery of acquired immunological tolerance

4. THE JOURNAL OF EXPERIMENTAL BIOLOGY 207 (23): 4013-4014 THE BIRTH OF TRANSPLANTATION IMMUNOLOGY: THE BILLINGHAM–MEDAWAR EXPERIMENTS AT BIRMINGHAM UNIVERSITY AND UNIVERSITY COLLEGE LONDON There are very few scientific papers that continue to have an impact on research more than 50 years after their initial publication. A paper published in J. Exp. Biol. in 1951 by Rupert Billingham and Peter Medawar is one such paper. (The technique of free skin grafting in mammals)

5. Evidence for active host defense against cancer • 80 years of immunotherapy. (Currie GA, 1972, Br. J. Cancer 26: 141) • A critique of the evidence for active host defense against cancer, based on personal studies of 27 murine tumors of spontaneous origin. (Hewitt HB, et al. 1976, Br. J. Cancer 33:241)

6. Circumvent Evidence • The high frequency of cancers in immunosuppressed patients • Extremes of age • Immunosuppressive drugs • Tumors that are infiltrated by T cells have an improved prognosis • Spontaneous regression occurs • Melanoma, breast, lung cancers, etc • Circulating tumor antibodies • MHC-I down-regulation

7. The ways to prove effective tumor immunity： 7

8. Immune surveillance, animal model

9. Full immunity needed to fight cancer   CD4+ T cells contribute to the remodeling of the microenvironment required for sustained tumor regression upon oncogene inactivation  (Cancer Cell, 2010, 18:485-498)

10. Full immunity needed to fight cancer

11. A fight between immune cells and cancer But, sometimes we lose

12. THE GREAT ESCAPE: IMMUNE EVASION VERSUS TUMOR PROGRESSION 12

13. 有些事只有在變動的狀況才看得見事物的本質

14. Difficulties due to study model

15. Tumor formation of Ras-over-expression cells in BALB/c It is a dynamic process death survival

16. Immune selection in the development of cancer: no two tumors are alike 16 Fey MF & Tobler A 1996 • Immune escape and unchecked proliferation • Microevolution, selection of immune resistance • Initiation, proliferation diversification

17. Use of tumor cell lines • Commonly derived from advanced tumors • Retain the genetic instability and lose the ability of adaptations • introduced in large numbers (more than 106 cells)

18. Successful immunization for cancer 18

19. Immune therapy (T cell-based) 19 • Recombinant and synthetic vaccination • Cytokine treatments (IL-2; GM-CSF; IFN) • Cellular therapy with tumor-specific CTL • Engineered macrophages (Th1…?) • Antigen-pulsed macrophages or dendritic cells

20. Tumor associated antigens • Tumor-specific shared antigens: restricted in expression to tumors and immune privilege sites. • Tissue-specific differentiation antigens: tyrosinase. (melanoma) • Tumor-specific antigens: mutated, tranlocated genes. • Ubiquitous antigens with over-expression in tumors. Rosenberg SA. 1999, Immunity 10:281

21. Peptide epitopes for melanoma Nicholaas P, et al. 1999, Curr Opin Oncol 11:50

22. Tumor and activated T cells Two major pathways for TCL: Fas-mediated and perforrin-mediated

23. HLA class I molecules 23 Antigen presentation for T cells, cytotoxic Inhibitory signals for NK cells

24. Immune escape/erosion • Making invisible • Resistance to death • Repelling • Active suppression • ….

25. An overview

26. Examples • Down-regulation of the class I presentation pathway • Resistance to killing • Antigen-specific mechanisms (Treg?) • Global mechanisms

27. Antigen-specific mechanisms • Tumor antigen-loss variants Loss of the melanoma tumor-associated antigen in patients with recurrent metastatic melanoma (J Clin Invest 1996, 98:1633) • Tolerance B cell tumors expressing class II induced a rapid tolerance of cognate CD4 T cell carrying a transgenic TCR. (PNAS, 1998, 95:1178)

28. Down regulation of the MHC class presentation pathway • Downregulation of MHC class I expression is frequently seen in human tumors. • Loss of MHC-I as a mechanism for tumor escape from CTL-mediated elimination (longitudinal study of melanoma patients) • Five major HLA altered phenotypes found in tumor tissues (Human Immunol. 2000, 61:65)

29. The five altered phenotypes NormalA1A2B8B35Cw7Cw4 1. Total loss- 2. Haplotype lossA1B8Cw7 3. Locus lossA1A2B8B35 4. Allelic lossA2B8B35Cw7Cw4 5. Compound phenotypeA1 (Human Immunol. 2000, 61:65)

30. Resistance to killing • Defective Fas pathway • Resistance to Granzyme B Cytotoxic T cells Innate immunity • Fas-L and Neutrophil

31. Nature 407, 789 5(2000) Fas signal 31 Apaf-1 Caspase 9 Caspase 3 Death substrates Apoptosis

32. Key person in Fas signal study Peter H. Krammer Shigekazu Nagata For a man who spends most of his time thinking about death, Shigekazu Nagata is remarkably upbeat. For over a decade, he has been making a name for himself with research on apoptosis, the mechanism of programmed cell death, and during that time he has watched the field come alive. Nature Medicine  7, 759 (2001) BC Yang 2008/2/20

33. Loss sensitivity to Fas-mediated apoptosis • FLICE-like inhibitory proteins • Bcl-2, Bcl-XL • defected sphingomyelinase activation (ceramide) • decoy receptor 3 (DcR3), soluble Fas

34. Fas-L+-melanoma cells are relatively resistant to killing of neutrophils B16F10 Chen YL, et al J. Immunol. 171:1183-1192.

35. Global Mechanisms • TGF-beta • IL-10 • Growth in immune privilege sites • Mucin production: interfering intercellular adhesion • Fas-L? (Fas counterattack) • Tumor extracellular matrix

36. TGF-b signaling in tumor signaling and cancer progression

37. IL-10 (Th3 or Treg) Tumors or other cells in environments

38. Immune cells are there! But, in peripheral stroma area (cell cuffing) Shaping T cell functions by extracellular matrix. NPC Done by 張逸如 (1998) Hematoxylin/eosin staining CD3+ cells

39. How can tumor cells highjack immune cells?Extracellular matrix shapes Fas-mediated apoptosis. • Protective: U-118, Huh-7, HeLa, A549, NIH3T3 • Non-protective: MCF-7, HepG2 Su CC, et al. 2007 Phosphatidylinositol 3-Kinase/Akt activation by integrin-tumor matrix interaction suppresses Fas-mediated apoptosis in T cells. J Immunol 179:4589-4597.

40. Coculture with tumor cells inhibits the Fas-mediated apoptosis.

41. Direct cell contact is required. NCKU

42. Apoptosis program is inhibited. Caspase 3 Apaf-1 Caspase 9 Death substrates Apoptosis NCKU

43. Fas-mediated death is not suppressed by components of MAPK, NFkB, PKA, HSP70.

44. What will happen when Fas-stimulated immune cells resist to die? Tissue environment ? Tumor Fas-L Immune cells Fas Cytokines ?

45. T cells in tumor microenvironment Jurkat and Molt-4 cells were cultured alone (lane 1) or in the presence of U118(V), U118(R), U373(V), or U373(R) (lanes 2, 3, 4, and 5, respectively) for 24 h. 林亨楷 J Immunol, 2003, 171: 3947–3954.

46. Regulatory T cells: the third man Shimon Sakaguchi (2000) Regulatory T Cells Key Controllers of Immunologic Self-Tolerance Cell 101: 455-458

47. A very dynamic interactions • Tumor microenviroment • Individual properties

48. A FasL mystery • Tissue dependant • Reverse signaling? • Other than death-triggering

49. Joe O’connell, 2002, Fas ligand and the fate of antitumorur cytotoxic T lymphocytes. Immunology 105:263-266.

50. In vivo consequences of Fas-L expression by tumors(using over-expression system) • Enhanced rejection • Delayed rejection CT26# B16-F10 Renca, MH134, L5178Y, B16-BL6, CT26* Note: *:syngenic, nude, SCID #: allogenic, lpr Ref: Lejeune FJ et al., 1998, Curr. Op. Immunol.